Regulatory Mochanisms For Proliferation, Differentiation, And Apoptosis of Leukemic Cells In Reference To Cell Cycle Phases.

白血病细胞增殖、分化和凋亡与细胞周期阶段相关的调节机制。

• 批准号: 06454349
• 负责人: SAITO Masaki
• 金额: $ 4.67万
• 依托单位: Hokkaido University (1995)Jichi Medical University (1994)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454349/
• 关键词: Retinoblastoma gene product (Rb); Extracellular Matrix (ECM) glycoproteins; Tenascin-C; Apoptosis inhibitory gene bcl-2; Differentiation expression; Cell cycle; Apoptosis susceptibility; Stromal-epithelial interactions; 細胞分化の制御; アポトーシス抑制遺伝子bcl-2

During the period from 1994 to 1995, the following new findings were obtained, and most of them were presented in some of the international conferences. and published in the specialized journals : (1) The inhibitory activity of TGF-beta on the growth of human monoblastic cell line JOSK-I cells was observed at the late G1 phase, but not at the S or G2/M phase of cell cycle. TGF-beta did not affect the level of both mRNA and protein of cdc2 gene, but did inhibit the translation rate of cdc2 gene and the cdc2 kinase activity. Thus, the growth inhibition by TGF-beta was closely related to the inhibition of RB protein (Rb) phosphorylation (the significant storage of unphosphorylated Rb) followed by the inhibition of DNA synthesis, and the unphosphorylated Rb storage was suggested to occur through the inhibition of cdc2 kinase activity by TGF-beta. On the other hand, the growth inhibitory activity of aphidicoline was observed at the G1/S phase with little unphosphorylated Rb. The differentia … More tion-inducer, phorbol ester TPA,also did unphosphorylate Rb protein, which was suggested to occur in the differentiation induction followed by the cell cycle arrest. With all these data together, it was suggested that some cdc2 gene products or an unidentified Rb kinase might be intimately involved in the Rb phosphorylation. (2) The experimental model system was established, in which, using human myelogenous leukemia cell line HL-60 cells, the differentiation induction (the differentiation commitment and the expression of differentiation properties) and the apoptosis induction could be separately analyzed. The results obtained were as follows : the NBT reducing acitivity and the apoptosis induction were independently observed, and the expression of both myc and bcl-2 genes was inhibited 6 hrs after the initiation of differentiation induction. During the period of 24-36 hrs after the initiation of differentiation induction, all the events necessary for the apoptosis induction occurred closely relating to the inhibition of bcl-2 expression. (3) The biological function of the extracellular matrix (ECM) glycoprotein, tenascin, was investigated since ECM glycoproteins in general have recently become clarified to play an important role in the regulation of cell proliferation and differentiation through the cell-to-cell adhesion and recognition. The expression of tenascin-C was termporally and spatially regulated in the strict manner at the stromal-epithelial interactions observed in tumor formations. Although the physiological regulators for tenascin-C expression-induction are still unknown, the do novo synthesis of tenascin-C in the stromal-epithelial interactions was induced with the soluble epithelial growth factor (EGF), and this induction was inhibited with steroid hormons such as hydrocortisone. Some stromal cell lines, which were deficient in the tenascin-C expression, could be first established from the bone marrows of tenascin-C gene knock-out mice, and then, usi Less

在1994年至1995年期间，获得了以下新的调查结果，其中大部分在一些国际会议上作了介绍。并发表在专业期刊上：(1)转化生长因子-β对人单核细胞株JOSK-I细胞生长的抑制作用出现在细胞周期的G1期晚期，而在细胞周期的S或G2/M期则未观察到。转化生长因子-β对cdc2基因的mRNA和蛋白水平均无影响，但对cdc2基因的翻译率和cdc2激酶活性均有抑制作用。因此，转化生长因子-β抑制细胞生长与抑制Rb蛋白(Rb)的磷酸化(未磷酸化的Rb显著储存)进而抑制DNA合成密切相关，而非磷酸化的Rb的储存可能是通过抑制cdc2的活性来实现的。另一方面，在G1/S期观察到除草剂对细胞生长的抑制作用，Rb几乎没有被磷酸化。The Difference…更多的诱导剂佛波酯TPA也使Rb蛋白去磷酸化，这可能发生在分化诱导和细胞周期停滞的过程中。所有这些数据表明，某些cdc2基因产物或一个未知的Rb激酶可能与Rb的磷酸化密切相关。(2)建立了以人髓系白血病细胞株HL-60为研究对象的实验模型体系，分别分析了分化诱导(分化承诺和分化特性的表达)和凋亡诱导。结果表明：诱导分化6h后，NBT还原活性和细胞凋亡诱导活性独立存在，myc和bcl2基因的表达均受到抑制。在分化诱导开始后的24-36小时内，所有诱导凋亡所必需的事件都与bcl2表达的抑制密切相关。(3)由于细胞外基质(ECM)糖蛋白通过细胞与细胞间的黏附和识别在调节细胞增殖和分化中发挥重要作用，因此对ECM糖蛋白的生物学功能进行了研究。在肿瘤形成中观察到的间质-上皮相互作用中，Tenascin-C的表达受到严格的时间和空间调控。尽管诱导Tenascin-C表达的生理调节因子尚不清楚，但在基质-上皮相互作用中，Tenascin-C的从头合成是由可溶性上皮生长因子(EGF)诱导的，这种诱导作用可被氢化可的松等类固醇激素抑制。从Tenascin-C基因敲除小鼠的骨髓中可以建立一些Tenascin-C表达缺失的基质细胞系，然后再用更少的

项目成果

Kizaki M,Nakajima H,Mori S,Koike T,Morikawa M,Ohta M,Saito M,Koeffler HP,Ikeda Y: "Novel Retinoic Acid,9-Cis Retinoic Acid,in Combination With AII-Trans Retinoic Acid Is an Effective Inducer of Differentiation of Retinoic Acid-Resistant HL-60 Cells." Bloo

Kizaki M、Nakajima H、Mori S、Koike T、Morikawa M、Ohta M、Saito M、Koeffler HP、Ikeda Y：“新型视黄酸，9-顺式视黄酸，与 AII-反式视黄酸结合是一种有效的诱导剂

斎藤政樹: "エイジング科学-21世紀の老化研究(積田享編)(現代化学増刊)" 東京化学同人(東京), 28 (1994)

齐藤正树：“衰老科学 - 21世纪的衰老研究（须木田章编辑）（现代科学特别版）”东京化学同人（东京），28（1994）

Kizaki, M., Saito, M., et al.: "Novel Retinoic Acid.9-Cis Retinoic Acid,in Combination With All-Trans Retinoic Acid Is an Effective Inducer of Differentiation ---" Blood. 83. 3289-3297 (1994)

Kizaki, M.、Saito, M. 等人：“新型视黄酸。9​​-顺式视黄酸与全反式视黄酸结合是分化的有效诱导剂 ---”血液。

Tsunoda, A., Saito, M., et al.: "IL-3-associated Expression of Gangliosides in Mouse Myelogenous Leukemia NFS60 Cells Introduced with IL-3 Gene : Expression of Ganglioside GD1a and A Key Involvement of CMP-NeuAc : Lactosylceramide alpha 2,6-Sialyltransfer

Tsunoda, A., Saito, M. 等人：“引入 IL-3 基因的小鼠骨髓性白血病 NFS60 细胞中神经节苷脂的 IL-3 相关表达：神经节苷脂 GD1a 的表达和 CMP-NeuAc 的关键参与：乳糖神经酰胺

Kojima N,Saito M,Tsuji S: "Role of Cell Surface O-Linked Oligosaccharides in Adhesion of HL60 Cells to Fibronectin:Regulation of Integrin-Dependent Cell Adhesion by O-Linked Oligosaccharide Elongation." Exp.Cell Res.214. 537-542 (1994)

Kojima N、Saito M、Tsuji S：“细胞表面 O 连接寡糖在 HL60 细胞与纤连蛋白粘附中的作用：通过 O 连接寡糖延伸调节整合素依赖性细胞粘附。”